Vio® Dyes technical specifications
Bright dyes for multicolor flow cytometry
Fluorochrome-conjugated MACS® Antibodies are perfectly
suited for identification and enumeration of human, mouse,
rat, or non-human primate cells.
Visit www.macsantibodies.com for a complete product
list of MACS Antibodies, Detection Kits, Cytokine Secretion
Assays, and Control Cocktails.
Vio® Dyes represent a family of fluorochromes for flow
cytometry and fluorescence microscopy developed
by Miltenyi Biotec. VioBlue®, VioGreen™, VioBright™ FITC,
PE-Vio® 615, PE-Vio® 770, PerCP-Vio® 700, Vio® 515, VioBright™
515, APC-Vio® 770 are characterized by high fluorescence
intensities and low spillover, making them an ideal choice
for multicolor applications. Combined with traditional
fluorochromes, such as FITC, PE, PerCP, and APC, the new
Vio Dyes expand Miltenyi Biotec’s antibody offering and
allow researchers a greater selection of antibodies for
multiparameter cell analysis.
VioBlue
VioGreen
VioBright 515 /
Vio 515
VioBright FITC
PerCP-Vio 700
PE-Vio 770
PE-Vio 615
APC-Vio 770
450
Violet laser
(405nm)
500
Blue laser
(488nm)
550
Yellow laser
(561 nm)
600
650
Red laser
(635 nm)
700
750
800
Wavelength (nm)
Figure 1: Emission spectra of Vio Dyes. Emission spectra of Vio Dyes: VioBlue, VioBright 515, Vio 515, VioGreen, VioBright FITC, PE-Vio 615, PerCP-Vio
700, PE-Vio 770 and APC-Vio 770.
Vio® Dyes technical specifications | April 20171/18
Overview of flurochromes available from Miltenyi Biotec
Excitation
laser (nm)
Exmax (nm)
VioBlue
405
400
452
V1
450/50
V1
450/50
VioGreen
405
388
520
V2
525/50
V2
525/50
Fluorochrome
Emmax (nm)
MACSQuant Analyzer
Channel
Filter (nm)
MACSQuant VYB
Channel
Filter (nm)
VioBright 515
488
488
514
B1
525/50
B1
525/50
Vio 515
488
488
514
B1
525/50
B1
525/50
VioBright FITC
488
496
522
B1
525/50
B1
525/50
FITC
488
495
520
B1
525/50
B1
525/50
PE
488 or 561
565
578
B2
585/40
Y1
586/15
PE-Vio 615
488 or 561
565
619
B3
655–730
Y2
615/20
PerCP
488
482
675
B3
655–730
N/A
N/A
PerCP-Vio 700
488
482
704
B3
655–730
N/A
N/A
PE-Vio 770
488 or 561
565
775
B4
750 LP
Y4
750 LP
APC
561 or 635
652
660
R1
655–730
Y3
661/20
APC-Vio 770
561 or 635
652
775
R2
750 LP
Y4
750 LP
Table 1: Fluorochrome specifications.
Miltenyi Biotec dyes
Other dyes
VioBlue®
Alexa Fluor® 405, BD™ Horizon™, V450, BV 421™, Calcein Violet 450 AM, Cascade Blue®, DAPI, Vybrant®,
DyeCycle™ Violet, eBFP, eFluor® 450, Hoechst Dyes, Pacific Blue™, Zombie Violet™
VioGreen™
Alexa Fluor® 430, AmCyan, BD™ Horizon™ V500, BV510™, Cascade Yellow™, Krome Orange™,
Pacific Orange™, Qdot® 525 Zombie Aqua™
FITC, VioBright™ FITC, Vio® 515,
VioBright™ 515
Alexa Fluor® 488, Calcein AM, DyLight® 488, CFSE, GFP, SYTOX® Green, Vybrant® DyeCycle™ Green, YFP,
Zombie Green™, BD Horizon™ Brilliant Blue 515
PE
Cy™3, Vybrant® DyeCycle™ Orange
PerCP, PerCP-Vio® 700
PE-Cy™5, PE-Cy™5.5, Per-Cy™5.5, Per-CP-eFluor® 710, Propidium iodide, 7-AAD
PE-Vio® 615
ECD, PE-Texas Red®, PE-CF594, PE/Dazzle™ 594, PE-eFluor® 610
PE-Vio® 770
PE-Alexa Fluor® 750, PE-Cy™ 7
APC
Alexa Fluor® 647, Alexa Fluor® 700, APC-Alexa Fluor® 700, Cy™5, DRAQ5®, eFluor® 660
APC-Vio® 770
APC-Alexa Fluor® 750, APC-Cy™7, APC-eFluor™ 780, APC-H7, Zombie NIR™
Table 2: Available Vio Dye fluorochromes.
Support Material
•Fluorescence spectra overview poster
•Fluorochromes and instruments poster
•Fluorochrome brightness index
•Lot-to-lot consistency data
•Staining enhancement products
• Viability dyes
Vio® Dyes technical specifications | April 20172/18
VioBlue® and VioGreen™ Fluorochromes are a novel
range of violet laser excited dyes developed exclusively
by Miltenyi Biotec. Designed to maximize the potential
of a flow cytometer’s violet laser, they not only show
superior optical performance compared with many
other 405 nm excited fluorochromes, but also significant
advances in brightness, signal-to-noise ratios, and intralaser compensation requirements. In addition, neither
fluorochrome exhibits significant levels of photoinduced
degradation, and can consequently be used for many
different applications, such as fluorescent microscopy.
A
Intensity
Violet laser dyes
330
VioBlue® Dye
Fluorochrome
Absorption
max. (nm)
Emission
max (nm)
VioBlue
400
455
Pacific Blue
401
452
Cascade Blue
401
419
Alexa Fluor 405
401
421
eFluor 450
405
450
BD Horizon V450
404
448
BD Horizon Brilliant™ Violet 421
407
421
Table 3: Absorption and emission maximums of fluorochromes related
to VioBlue.
Laser and filter compatibility
With a maximum absorption and emission at 400 nm and 455
nm respectively, VioBlue Fluorochromes are fully compatible
with standard filter sets from all major flow cytometry
hardware providers, giving researchers the flexibility to use
the VioBlue Dye with existing laboratory platforms (figure 2).
Pacific Blue
Cascade Blue
Alexa Fluor 405
VioBlue
B
Intensity
•Coumarin-based dye with excitation and emission
wavelengths of 405 nm and 455 nm, respectively
•Superior alternative to Pacific Blue, Alexa Fluor 405,
or BD Horizon V450
•Multiplexing of VioBlue with other fluorochromes
is easily possible, adding to the variety of marker
combinations for multiparameter cell analysis
• Combined use with the VioGreen Dye
430
380
Wavelength (nm)
380
430
480
530
Wavelength (nm)
Pacific Blue
Cascade Blue
Alexa Fluor 405
VioBlue
Figure 2: Absorption (A) and emission (B) spectra of VioBlue
compared to Pacific Blue, Cascade Blue, and Alexa Fluor 405.
The blue box represents the 450/50 nm filter.
Enhanced brightness
Human peripheral blood mononuclear cells (PBMC) and mouse
splenocyte (MS) cells stained with VioBlue® exhibit excellent
discrimination between positively and negatively stained
populations over a wide range of antibody specificities
(figure 3). Furthermore, VioBlue provides a superior alternative
to many spectrally similar conjugates for the V1 channel,
further ­increasing the options of multicolor analysis (figure 4).
Vio® Dyes technical specifications | April 20173/18
750
750
Side scatter
B 1000
Side scatter
A 1000
500
250
0
-1 0 1
10¹
10²
High stability during fixation
It is of crucial importance for a conjugate to retain its
fluorescent properties after fixation, in order to allow
researchers to maximize the use of biological samples.
The VioBlue Dye has a very high stability after fixation with
paraformaldehyde (figure 5), equal to or exceeding many
other spectrally similar fluorochromes.
500
250
0
-1 0 1
10³
CD20-VioBlue
10¹
10²
10³
CD123-VioBlue
A
C 1000
D 1000
750
750
B
10³
10³
10¹
10²
CD8-VioBlue
10³
250
0
-1 0 1
10¹
10²
10³
VioBlue
VioBlue
PacificPacific
Blue Blue
Relative cell number
Relative cell number
CD303-PE
-1 0 1
10¹
10²
10³
1
0
-1
CD123-VioBlue
-1 0 1
10¹
10²
10³
CD123-VioBlue
CD90.2-VioBlue
Figure 3: Human peripheral blood mononuclear cells (PBMC) or mouse
splenocyte (MS) cells were stained with CD20-VioBlue (A; PBMCs), the
rare cell marker CD123‑VioBlue (B; PBMCs), CD8-VioBlue (C; PBMCs),
or CD90.2-VioBlue (D; MS) and analyzed by flow cytometry using the
MACSQuant Analyzer.
-1 0-1 10 1 10¹ 10¹10² 10² 10³ 10³
VioBlue
1
0
-1
10¹
Figure 5: CD123-VioBlue staining before (A) and after (B) fixation with
3.7% paraformaldehyde, indicating a minimal decrease in fluorescence
after fixation.
Lot-to-lot variation
Reproducible lot-to-lot antibody performance ensures the
integrity of long-term studies, and is crucial in comparing
experimental results over a period of time. All MACS®
Antibodies are vigorously tested to ensure consistent
quality, thus eliminating the need for repeat testing
arising from reagent degradation or failure (figure 6).
A
Pacific Blue
Figure 4: Fluorescence intensity of cells labeled with CD14 antibodies
conjugated to either VioBlue or Pacific Blue.
-1 -10 01 1 10¹10¹ 10²10² 10³10³
-1 -10 01 1 10¹10¹ 10²10² 10³10³
CD3-VioBlue
CD3-VioBlue
CD4-VioBlue
CD4-VioBlue
C
D
Relative cell number
Relative cell number
Decreased spillover
VioBlue® Dyes exhibit minimal spillover into the V2 channel,
making them perfect candidates for multicolor panels,
which utilize both violet channels. Furthermore, VioBlue
is negligibly excited by the 488 nm laser, and thus requires
no compensation between the V1 and B1 channels.
B
Relative cell number
Relative cell number
0
-1 0 1
10¹
Relative cell number
Relative cell number
250
500
10²
Relative cell number
Relative cell number
500
CD303-PE
Side scatter
Side scatter
10²
-1 -10 01 1 10¹10¹ 10²10² 10³10³
-1 -10 01 1 10¹10¹ 10²10² 10³10³
CD20-VioBlue
CD20-VioBlue
CD45-VioBlue
CD45-VioBlue
Figure 6: Lot-to-lot variation for different lots of CD3 (A), CD4 (B), CD20
(C), and CD45 (D) antibodies conjugated to VioBlue. Black and blue lines
represent different antibody lots.
Vio® Dyes technical specifications | April 20174/18
Emission
max. (nm)
VioGreen
388
520
Pacific Orange
400
551
Krome Orange
398
528
AmCyan
457
491
Horizon V500
415
500
BD Horizon Brilliant™ 510
405
510
Table 4: Absorption and emission maximums of fluorochromes
comparable to comparable to VioGreen™.
Laser and filter compatibility
With a standard 525/50 filter set, the VioGreen™ Dye exhibits
a better spectral profile than Pacific Orange or AmCyan
(figure 7).
A 1000
B 1000
750
750
Side scatter
Absorption
max. (nm)
500
250
0
-1 0 1
10¹
10²
500
250
0
-1 0 1
10³
CD14-VioGreen
C 1000
D 1000
750
750
500
250
0
-1 0 1
10¹
10²
10¹
10²
10³
CD20-VioGreen
Side scatter
Fluorochrome
Side scatter
•Large Stokes shift fluorochrome, emitting strong
fluorescence at 520 nm upon excitation at 405 nm
•Significantly increased mean fluorescence intensities
and higher stain indices over Pacific Orange,
Krome Orange, and BD Horizon V500
• Non-protein fluorochrome
• Suitable for ethanol fixation
• Combined use with the VioBlue® Dye
•Perfectly suited for multiparameter cell analysis
using all current flow cytometers
Enhanced brightness
Like most dyes designed for the violet laser, VioGreen
shows a lower fluorescence intensity compared to other
well-established fluorochomes, such as PE. However,
specific human and mouse cellular populations are easily
distinguishable between positively and negatively labeled
cells (figure 8). In addition, many VioGreen Dyes exhibit
brighter fluorescence compared to spectrally similar
conjugates, such as Pacific Orange (figure 9 and table 5),
AmCyan, and Horizon V500, when measured by mean
fluorescence intensity (MFI) or stain index (normalized
signal-to-noise ratio).
Side scatter
VioGreen™ Dye
500
250
0
-1 0 1
10³
CD15-VioGreen
10¹
10²
10³
CD8a-VioGreen
A
300
350
400
450
VioBlue
Pacific Blue
Relative cell number
Intensity
Figure 8: Human peripheral blood mononuclear cells (PBMC) or
mouse splenocyte (MS) cells were stained with CD14-VioGreen (A;
PBMCs), CD20-VioGreen (B; PBMCs), CD15-VioGreen (C; PBMCs), or
CD8a-VioGreen (D; MS) and analyzed by flow cytometry using the
MACSQuant® Analyzer.
500
Wavelength (nm)
B
-1 0 1
Intensity
VioGreen
10¹
Horizon V500
10²
10³
Krome Orange
Figure 9: Analysis of human PBMCs using CD3 antibodies conjugated
to either VioGreen, Horizon V500, or Krome Orange.
419
519
619
694
Wavelength (nm)
VioGreen
AmCyan
Pacific Orange
Figure 7: Absorption and emission spectra of VioGreen.
Vio® Dyes technical specifications | April 20175/18
Conjugate
Clone
MFI
Stain
index
Percent
spillover
V1
B1
CD3-VioGreen
BW264/56
13
13
5.5
0.5
CD3-Horizon V500
UCHT1
11
9
8.0
1.5
CD3-Krome Orange
UCHT1
6
7
3.0
1.0
CD8-VioGreen
BW135/80
16
17
6.0
0.2
CD8-Pacific Orange
BW135/80
10
11
3.0
0.5
Table 5: MFI, stain indices, and % spillover of CD3-VioGreen,
CD3‑Horizon V500, CD3-Krome Orange, CD8-VioGreen, and
CD8-Pacific Orange.
High stability during paraformaldehyde
and ethanol fixation
VioGreen™ Dyes exhibit strong photostability during
fixation, with only very minimal photodegradation, thus
highlighting VioGreen’s suitability as a fluorochrome for
use in studies that require fixation (figure 10). It is also
suitable for ethanol fixation.
750
750
Side scatter
B 1000
Side scatter
A 1000
500
250
0
-1 0 1
10¹
10²
CD14-VioGreen
10³
500
Blue laser dyes
VioBright™ FITC, VioBright 515 and
Vio® 515 Dyes
VioBright™ FITC, VioBright 515 and Vio® 515 are revolutionary
blue laser (488 nm) excitable dyes, developed exclusively by
Miltenyi Biotec. The innovative VioBright technology allows
for an increased number of fluorochrome molecules per
antibody, as compared to conventional conjugation. This
results in dyes that emit a signal detected in the standard
FITC channel, but with brightness levels similar to PE. These
blue laser dyes expand the dimensions of multicolor flow
analysis and provide a bright alternative for confident
detection of rare cells, dim, and uncharacterized markers.
Dye
Features
Application
VioBright FITC
(1st generation
VioBright dye)
•
High brightness
Detection of
extracellular
markers
VioBright 515
(2nd generation
VioBright dye)
•
Very high brightness
•
Low spill over
•
High fixation- &
photostability
Detection of
extracellular
markers
Vio 515
(Organic dye)
•
Moderate brightness,
•
Low spill over
•
High fixation- &
photostability
Detection of
intracellular
markers
250
0
-1 0 1
10¹
10²
10³
Table 6: Features and application specifications for VioBright FITC,
VioBright 515 and Vio 515 in comparison.
CD14-VioGreen
Figure 10: CD14-VioGreen staining before (A) and after (B)
paraformaldehyde fixation, indicating a minimal decrease in
fluorescence after fixation.
Laser and filter compatibility
Excitable by the blue laser (488 nm), VioBright 515 and Vio 515
display peak excitation and emission at 488 nm and 514 nm,
respectively (figure 11 and table 7). VioBright FITC is excited
at 496 nm with a peak emission at 522 nm. The high intensity
fluorescent signals can be detected with a standard FITC filter
such as 525/50 on MACSQuant Instruments. Thus, no change in
the filter or cytometer is required. Along with PE, the VioBright
FITC, VioBright 515 and Vio 515 dyes enhance the potential of
the blue laser, one of the most common laser lines available for
single laser to multi-laser instruments.
Fluorochrome
Absorption
max. (nm)
Emission
max. (nm)
VioBright 515
488
Vio 515
488
514
VioBright FITC
496
522
FITC
495
520
Alexa Fluor 488
495
519
BD Horizon Brilliant Blue 515
490
515
514
Table 7: Absorption and emission maximums of fluorochromes
comparable to VioBright FITC, VioBright 515 and Vio 515.
Vio® Dyes technical specifications | April 20176/18
A
A
Absorption spectra
B
10³
1.000
10³
Absorption
0.600
10²
CD4-APC
CD4-APC
10²
0.800
10¹
1
0
-1
-1 0 1
0.400
10¹
10²
10¹
1
0
-1
-1 0 1
10³
CD25-PE
10¹
10²
10³
CD25-VioBright FITC
0.200
0.000
440
450
460
470
480
490
500
510
520
530
Wavelength (nm)
Figure 13: Human PBMCs were stained with CD25 antibodies
conjugated to PE or VioBright FITC, as well as with CD4-APC, and
analyzed by flow cytometry using the MACSQuant Analyzer.
Cell debris and dead cells were excluded from the analysis based
on scatter signals and propidium iodide fluorescence.
Emission spectra
B
1
B
A
CD3-VioBlue
Emission
0.8
0.6
0.4
0.2
0
500
510
520
530
540
550
560
570
Wavelength (nm)
VioBright FITC
VioBright 515
Figure 14: Human whole blood was stained with CD3-VioBlue® and
CD56-VioBright™ 515 or CD56-BB515, respectively.
Vio 515
FITC
Figure 11: Absorption (A) and emission (B) spectra of VioBright™ FITC,
VioBright 515 and Vio 515 compared to FITC.
Reliable analysis of rare cells requires specific identification
of low frequency cellular subset and clear target population
resolution. Thus, highly specific antibodies together with
bright conjugates are a prerequisite for detection of such
low frequency populations. As demonstrated in figures 11–14,
VioBright FITC and VioBright 515 represent bright dye
alternatives for optimal detection of your subpopulation of
interest and detailed phenotypic analysis.
A
CD56-FITC
B
10³
CD34-VioBlue
Enhanced brightness
VioBright™ FITC, VioBright 515 and Vio® 515 are superior
dyes for detection of low-expressed markers, as shown
in figure 12 and 13. With stain indices (SI) and mean
fluorescent intensities similar to PE, staining with these
novel dyes allows for an excellent resolution of positive
and negative populations.
10²
10¹
1
0
-1
-1 0 1
10¹
10²
CD133/1-PE
5
CD
6-
FI
TC
CD
B
-V
56
FI
TC
C
6
D5
B
-V
5
51
C
6
D5
-B
CD56-VioBright 515
CD56-BB515
580
15
B5
10³
CD34-VioBlue
490
10³
10²
10¹
1
0
-1
-1 0 1
10¹
10²
10³
CD133/1-VioBright FITC
Figure 15: CD34 + enriched human PBMCs were stained with CD133/1
antibodies conjugated to PE or VioBright FITC as well as with CD34VioBlue and analyzed by flow cytometry using the MACSQuant
Analyzer. Cell debris and dead cells were excluded from the analysis
based on scatter signals and propidium iodide fluorescence.
Figure 12: Human PBMCs were stained using CD56 antibodies
conjugated to FITC, VioBright FITC, VioBright 515, or BB515 and
analyzed by flow cytometry using the MACSQuant® 10 Analyzer.
Vio® Dyes technical specifications | April 20177/18
Conjugate
Stain index
VioBright FITC
PE
VioBright FITC
PE
3.3
2.0
5.0
4.0
CD335
4.8
4.0
8.5
11
CD133/1
3.5
4.7
4.7
4.7
CD25
Table 8: MFI and stain indices of CD25 (clone 4E3), CD335 (clone 9E2),
and CD133/1 (clone AC133) antibodies conjugated to either VioBright
FITC or PE.
Spill over
Compared to standard FITC, VioBright 515 and Vio® 515 dyes
display reduced spill over into the PE channel (table 8). In
contrast, VioBright FITC has a slightly higher spill over into
the PE channel. Thus, with nominal change in compensation
settings, these innovative dyes offer the benefit of a
brighter staining.
Fixation stability
Bright fluorochrome conjugates, such as PE and APC, are
often sensitive to fixatives like Methanol. For a successful
flow cytomeric analysis, which often involves staining
of intracellular markers, stability of the fluorochrome
conjugates is critical. VioBright 515 conjugates show excellent
stability to methanol- and paraformaldehyde-based fixatives
with only little compromise in brightness (figure 16).
35
30
Stain Index
MFI
25
20
15
10
5
0
MACSQuant
VYB
Compensation in
channel
Y1 (%)
MFI
Stain
index
66
106
8.5
0
A
CD4VioBright
FITC
A
CD4-FITC
29
49
6.5
0
A
CD4-PE
58
126
–
–
B
CD4VioBright
FITC
52
81
9.3
0
B
CD4-FITC
20
35
7.0
0
B
CD4-PE
39
73
–
–
Table 9: Compensation requirements in PE channel (B2) of CD4 (clone
VIT4) antibodies conjugated to VioBright FITC, FITC, or PE.
Conjugate
Clone
MFI
10.6
Stain
index
18.3
MACS Quant
Compensation
in Channel
B2 [%]
CD56-FITC
REA196
6.6
CD56-VioBright FITC
AF12
15.8
22.6
8.6
CD-VioBright 515
REA196
31.9
56.3
4.2
CD56-BB515
B159
4.9
4.4
6.2
Table 10: Mean fluorescent intensities (MFI) and stain indices of
anti-CD56 antibodies conjugated to FITC, VioBright 515, and BB515.
PFA Fixation
No Fixative
CD56(REA196)-VioBright 515
MethanolFixation
CD56(REA196)-PE
Figure 16: Human PBMCs were stained using CD56 antibodies (clone
REA196) conjugated to VioBright 515 and PE. In addition, cells were
analyzed before and after fixation using paraformaldehyde and 90%
methanol. Stained cells were analyzed by flow cytometry using the
MACSQuant® 10 Analyzer.
Photo stability
The stability of VioBright 515 upon exposure to confocal
laser light was analyzed at different time points on a Zeiss
LSM710 confocal microscope. Significantly higher mean
fluorescence intensity (MFI) compared to FITC conjugate,
were demonstrated for these time points (figure 17).
VioBright 515 shows comparable stability to Alexa Fluor 488
conjugated antibodies indicating the excellent suitability
for immunofluorescence microscopy.
% 100
Normalized Intensity
Sample Conjugate
MACSQuant
Analyzer 10
Compensation in
channel
B2 (%)
80
60
40
20
0
02:00
VioBright 515
07:00
FITC
12:00
Min
17:00
Alexa Fluor 488
Figure 17: Human PBMCs were stained with fluorochrome-conjugated
CD4 antibodies. Stained cell samples were fixed with p-formaldehyde
for 20 min and transferred to a 96-well plate for analysis. The plotted
curve is normalized over multiple time points.
Vio® Dyes technical specifications | April 20178/18
Blue laser tandem conjugates
A
PE-Vio® 615 Tandem Conjugate
1.0
PE-Vio® 615 is a tandem dye with PE as the donor dye and
Vio® 615 as the acceptor dye. This tandem dye is optimized
for efficient donor-to-acceptor dye energy transfer, high
fluorescent intensity, and low spillover into the donor dye
detection channel. Designed to be a superior alternative
to ECD, PE-Texas Red®, PE-efluor® 610, PE-CF594 and PE/
Dazzle™ 594, this Vio Dye expands the options for flexible
multicolor panel design and provides a bright dye for
confident detection of dim and rare markers.
0.8
Absorption max
(nm)
Emission
max (nm)
PE-Vio 615
565
619
ECD, PE-Texas Red
565
613
PE-efluor 610
565
606
PE-CF594
565
614
PE/Dazzle 594
566
612
Table 11: Absorption and emission maximums of fluorochromes
comparable to PE-Vio 615.
Laser and filter compatibility
The PE molecule shows peak absorption at two
wavelengths, 496 nm and 565 nm, respectively. This allows
for optimal excitation of PE-containing tandem dyes such
as PE-Vio 615 using blue, green, and yellow green laser lines
(488-561 nm). This enables the utilization of the maximum
potential of your instrument with flexible choice of laser
lines (figure 19). As depicted in figure 15, the extent of
absorption at 565 nm is greater than 496 nm nm and thus,
the maximum excitation of PE-Vio 615 can be achieved with
instruments such the MACSQuant VYB, which uses yellow
laser lines for excitation of PE and all PE containing tandem
dyes. The emission signal can be detected using typical
filters design to detect PE-Texas Red, such as 615/20 nm.
0.2
0
400
450
500
550
600
650
700
450
500
550
600
650
700
B
1.0
0.8
0.6
0.4
0.2
0
400
— PE-CF594 — PE-Vio 615 — ECD — PE-Dazzle 594
Figure 18: Absorption (A) and emission (B) spectra of PE-Vio 615 and
comparable fluorochromes.
A
B
10³
10³
10²
10²
CD3-FITC
Fluorochrome
0.4
CD3-FITC
•Optimal excitation with blue (488 nm),
green (532 nm), and yellow green (561 nm)
laser lines for maximum flexibility.
•Excellent brightness for confident detection of dim,
rare, and uncharacterized markers.
•Extensive portfolio of PE-Vio 615 dye conjugated to
recombinantly engineered REAfinity Antibodies for
higher reproducibility.
0.6
10¹
1
0
-1
-1 0 1
10¹
10²
CD4-PE-Vio 615
10³
10¹
1
0
-1
-1 0 1
10¹
10²
10³
CD4-PE-Vio 615
Figure 19: Human PBMCs were stained using CD4 antibodies (clone
VIT4) conjugated to PE-Vio615 followed by analysis using MACSQuant
VYB (A) and MACSQuant Analyzer (B).
Vio® Dyes technical specifications | April 20179/18
10³
CD335-VioBright FITC
10³
CD335-VioBright FITC
Enhanced brightness
PE-Vio® 615 is designed to offer a bright alternative to
comparable dyes such as PE-efluor 610, PE-CF594, ECD,
and PE/Dazzle 594 (figure 20). PE-Vio 615 not only allows for
high fluorescent intensities but also excellent separation of
positive population for higher stain indices (figure 21). This
property of PE-Vio 615 makes it an excellent dye for analysis
of dim and difficult to characterize markers (figure 22).
10²
10¹
1
0
-1
-1 0 1
10¹
10²
10³
10²
10¹
1
0
-1
-1 0 1
CD56-PE-Vio 615
— PE-eFluor610
— PE-CF594
— PE-Vio 615
— ECD
— PE-Dazzle 594
Figure 20: Human PBMCs were stained using CD4 antibodies
conjugated to PE-eFluor610, PE-CF594, PE-Vio615, ECD, PE-Dazzle594
and analyzed by flow cytometry using the MACSQuant VYB.
Conjugate
Clone
MFI
Stain
index
MACS Quant VYB
Compensation in
channel Y1 [%]
CD4-PEVio 615
Vit-4.3
236
309
3.0
CD4-ECD
SFCI12T4D11
133
185
3.3
CD4-PECF594
RPA-T4
201
295
2.6
CD4-PEeFluor610
RPA-T4
202
259
5.5
CD4-PE/
Dazzle594
RPA-T4
316
287
3.9
10²
10³
Figure 21: Human PBMCs were stained using CD56 antibodies (clone
REA196) conjugated to either PE-Vio 615 or PE and CD335-VioBright
FITC. Stained cells were then analyzed by flow cytometry using the
MACSQuant VYB.
Q1
25.2
Q2
1.03
Q1
25.1
Q2
1.09
Q4
73.6
Q3
0.13
Q4
73.5
Q3
0.31
Anti-IL-4-PE
CD4-PE-Vio 615
10¹
CD56-PE
Anti-IL-4-PE-Vio615
Figure 22: PBMCs were stimulated for 6 hours. After 2 hours, 1 µg/mL
BrefeldinA was added for the last 4 hours. Cells were surface stained
with CD4-FITC, fixed and permeabilized with the Inside Stain Kit
(Miltenyi Biotec) and stained intracellularly with CD154-VioBlue and
Anti-IL-4-PE or Anti-IL-4-PE-Vio615 (clone: 7A3-3). Cells were acquired on
a BD Fortessa. Cells are gated on CD4 + lymphocytes, numbers indicate
frequency among CD4 + cells.
Data courtesy: Petra Bacher, Clinic for Rheumatology and Clinical
Immunology, Charité – University Medicine Berlin, Berlin, Germany
Table 12: MFI and stain indices of CD4 antibodies conjugated to
PE-Vio 615, ECD, PE-CF594, PE-eFluor610, PE/Dazzle594.
Vio® Dyes technical specifications | April 201710/18
Fixation stability
Tandem conjugates are often sensitive to fixatives. Thus,
for a successful flow cytomeric analysis, stability of tandem
conjugates is critical. PE-Vio® 615 conjugate shows excellent
stability to paraformaldehyde-based fixative without any
increase in background signal (figure 23).
Without fixation
With fixation
10³
10¹
1
0
-1
-1 0 1
10¹
10²
10³
10²
10¹
1
0
-1
-1 0 1
CD56-PE
10¹
10²
10³
10¹
1
0
-1
-1 0 1
10¹
10²
10³
10¹
10²
10³
CD56-PE-Vio 615
10²
10¹
10²
10³
10¹
1
0
-1
-1 0 1
10¹
10²
10³
10²
10¹
t 24 h
1
0
-1
-1 0 1
CD56-PE-Vio 615
10¹
10²
10³
CD56-PE-Dazzle594
10¹
1
0
-1
-1 0 1
10¹
10²
10³
10²
10¹
1
0
-1
-1 0 1
CD56-PE-Dazzle594
10¹
10²
10³
CD56-PE-Vio 615
CD56-PE-Dazzle594
Compensation (%)
10²
Decrease in MFI (%)
10³
CD335-VioBright FITC
10³
t 0
1
0
-1
-1 0 1
10³
CD335-VioBright FITC
10²
10¹
10¹
CD56-PE-Dazzle594
10³
1
0
-1
-1 0 1
10²
CD56-PE-Vio 615
CD335-VioBright FITC
CD335-VioBright FITC
10²
CD56-PE
10³
CD335-VioBright FITC
10³
CD335-VioBright FITC
10²
10³
CD335-VioBright FITC
CD335-VioBright FITC
10³
CD335-VioBright FITC
Photo stability
The stability of PE-Vio 615 upon exposure to ambient
light (~850 Lux) was analyzed at different time points.
Significantly higher mean fluorescence intensities (MFI)
and stain indices (SI) for these time points, compared to
commercially available alternatives, were also demonstrated
(figure 23).
CD56-PE-Vio 615
CD56-PE-Dazzle594
CD56-PE-Dazzle594
Light exposing time (h)
without fixation
with fixation
Conjugate
Clone
MFI
Stain
index
MFI
Stain
index
CD56-PE
REA196
12.0
33.2
11.9
32.7
CD56-PEVio 615
REA196
39.9
70.9
40.1
71.4
CD56-PEDazzle594
HCD56
30.5
42.2
31.5
54.0
Light exposing time (h)
Figure 24: Human PBMCs were stained, using CD56 antibodies
conjugated to PE-Vio615 (clone REA196) or PE-Dazzle594 (clone HCD56).
Stained cells were then exposed to ambient light (~850 Lux) followed
by analysis at different time points by flow cytometry using the
MACSQuant VYB. Spill over in the Y1 channel, which is optimized for
detection of PE signal was also analyzed after exposure to light.
Figure 23: Human PBMCs were stained using CD56 antibodies (clone
REA196) conjugated to PE, PE-Vio615 or PE-Dazzle594. In addition, cells
were stained with CD335-VioBright FITC and analyzed before and after
fixation using paraformaldehyde. Stained cells were analyzed by flow
cytometry using the MACSQuant VYB.
Vio® Dyes technical specifications | April 201711/18
Fluorochrome
Absorption
max. (nm)
Emission
max. (nm)
PerCP
482
675
PerCP-Vio 700
482
704
PerCP-Cy5.5
490
695
A 1000
B 1000
750
750
Table 13: Absorption and emission maximums of fluorochromes
related to PerCP-Vio700.
Side scatter
•Combines the peridinin chlorophyll protein (PerCP)
and the new Vio® 700 dye
•Emits strong fluorescence at 655–730 upon
blue laser excitation at 488 nm
• Suitable for the B3 channel of the MACSQuant Analyzer
Enhanced brightness
Human peripheral blood mononuclear cells (PBMC)
and mouse spenocyte (MS) cells were stained with
PerCP-Vio 700, and consequently exhibited excellent
separation between positive and negative populations
over many different antibody specificities (figure 26).
Side scatter
PerCP-Vio® 700 Tandem Conjugate
500
250
0
-1 0 1
10¹
10²
500
250
0
-1 0 1
10³
CD20-PerCP-Vio 700
D 1000
750
750
Side scatter
C 1000
Side scatter
Laser and filter compatibility
Using a standard 655–730 bandpass filter, PerCP-Vio 700
exhibits a very narrow fluorescence spectral profile, thus
allowing the majority of light to be captured and retained
(figure 25).
500
250
0
-1 0 1
100
10¹
10²
10²
10³
500
250
0
-1 0 1
10³
CD45-PerCP-Vio 700
80
10¹
CD4-PerCP-Vio 700
10¹
10²
10³
CD90.2-PerCP-Vio 700
Intensity
70
Figure 26: Human peripheral blood mononuclear cells (PBMC) or
mouse spenocyte (MS) cells were stained with CD20-PerCP-Vio 700
(A; PBMCs), CD4-PerCP-Vio700 (B; PBMCs), CD45-PerCP-Vio 700
(C; PBMCs) or CD90.2-PerCP-Vio 700 (D; MS) and analyzed by flow
cytometry using the MACSQuant Analyzer.
60
50
40
30
20
10
0
350
500
650
800
High stability during fixation
PerCP-Vio 700 shows excellent fixation stabilities, with only
minimal decreases in fluorescence after fixation (figure 27).
Wavelength (nm)
B 1000
750
750
Figure 25: Absorption and emission spectra of PerCP-Vio 700.
The blue box represents the 655–730 filter.
Side scatter
A 1000
Emisson
Side scatter
Absorption
500
250
0
-1 0 1
10¹
10²
CD8-PerCP-Vio 700
10³
500
250
0
-1 0 1
10¹
10²
10³
CD8-PerCP-Vio 700
Figure 27: CD8-PerCP-Vio 700 before (A) and after (B) fixation with
3.7% paraformaldehyde, indicating a minimal decrease in fluorescence
after fixation.
Vio® Dyes technical specifications | April 201712/18
Photo-induced conjugate degradation
Analysis of the photo-induced degradation of CD14-PerCPVio 700 indicated no discernable changes after up to four
hours of continuous exposure to ambient light (~850 Lux).
Significantly higher mean fluorescence intensities (MFI)
and stain indices (SI) for these time points, compare to
commercially available alternatives (BD Biosciences),
were also demonstrated (figure 28).
t 0 min
CD14-PerCP-Cy5.5
CD14-PerCP
CD14-PerCP-Vio 700
t 120 min
t 240 min
CD11b-FITC
Marker
Clone
CD14-PerCP
TÜK4
53
44
36
CD14-PerCP-Vio 700
TÜK4
75
75
75
CD14-PerCP-Cy5.5
61D3
36
31
28
t 0 min
Stain index
t 120 min t 240 min
Figure 28: Photo-induced conjugate degradation of PerCP, PerCP‑Vio
700, and PerCP-Cy5.5 with corresponding MFI and SI. Conjugates
were exposed to ambient light for up to 4 hours, with negligible
degradation rates.
Vio® Dyes technical specifications | April 201713/18
Absorption
max. (nm)
750
750
PE-Vio 770
496, 565
775
496, 565
785
PE-Alexa Fluor 750
496, 546
775
Table 14: Absorption and emission maximums of fluorochromes
comparable to PE-Vio770.
250
10²
500
250
0
-1 0 1
10³
CD3-PE-Vio 770
D 1000
750
750
500
250
0
-1 0 1
10¹
10²
10¹
10²
10³
CD14-PE-Vio 770
C 1000
10³
CD19-PE-Vio 770
500
250
0
-1 0 1
10¹
10²
10³
CD4-Biotin/
Anti-Biotin-PE-Vio 770
Figure 30: Human peripheral blood mononuclear cells (PBMC) or
mouse splenocyte (MS) cells were stained with CD3-PE-Vio 770 (A;
PBMCs), CD14-PE-Vio 770 (B; PBMCs), CD19-PE-Vio770 (C; PBMCs), or
CD4-Biotin/Anti-Biotin-PE-Vio 770 (D; MS) and analyzed by flow
cytometry using the MACSQuant Analyzer.
100
80
70
Intensity
10¹
PE-Vio770
PE-Vio770
PE-Cy7
PE-Cy7
Relative cell number
Relative cell number
Laser and filter compatibility
The tandem dyes PE-Vio 770, PE-Cy7, and PE-Alexa Fluor 750
all use PE as the donor molecule, showing absorption at
496 nm and to a greater extent at 565 nm. Consequently,
flow cytometers, such as the MACSQuant VYB, equipped
with a yellow 561 nm laser, provide maximum excitation
to PE molecules, which in turn transfer this energy to
the respective acceptor molecule. Few differences in the
emission properties can be found between Vio 770, Cy7, and
Alexa Fluor 750, making each tandem best suited to flow
cytometry platforms using 750 LP filters (figure 21).
500
0
-1 0 1
Emission
max. (nm)
PE-Cy7
Side scatter
B 1000
Side scatter
Fluorochrome
A 1000
Side scatter
The PE-Vio® 770 Dye is a tandem conjugate, like PE-Cy™7,
that exploits the principle of fluorescence-resonanceenergy-transfer (FRET) allowing for large Stokes shifts between
the absorbed energy of a fluorescence donor (PE) and the
emission wavelength of a suitable acceptor (Vio 770) dye.
•Excitation with the blue (488 nm) or yellow (561 nm)
laser, emission in the near-infrared region at 775 nm
•The combination of Vio 770 as the acceptor dye and an
optimized chemistry have furnished a tandem dye that
is characterized by a high fluorescence intensity,
minimal spillover to adjacent detection channels, and
low non-specific binding to non-target cells to meet the
complexity of multiparameter cell analysis
•Higher MFI and stain index values, in addition to lower
compensation settings when compared to PE-Cy7
Enhanced brightness
PE-Vio 770 provides the greatest fluorescence intensity of
the Vio Dye family, with excellent separation of positively
and negatively stained cells (figure 30). When compared
to other spectrally similar tandem conjugates, such as
PE-Cy7 (figure 31) or PE-Alexa Fluor 750, PE-Vio 770 exhibits
significantly higher mean fluorescence intensities (MFI)
and stain indices, and requires less compensation (table 14).
These properties make PE-Vio 770 a far superior tandem
conjugate compared to PE-Cy7 or PE-Alexa Fluor 750.
Side scatter
PE-Vio® 770 Tandem Conjugate
60
-1 -1
0 10 1 10¹ 10¹ 10² 10² 10³ 10³
50
40
PE-Vio 770
30
20
10
0
400
450
500
550
600
650
700
750
800
850
PE-Cy7
Figure 31: Analysis of human PBMCs using CD8 antibodies (clone:
BW135/80) conjugated to either PE-Vio 770 or PE-Cy7. Concurrent
staining with CD14-PerCP and CD56-PE was performed to exclude
CD14 + and CD56+ cells from the analysis.
Wavelength (nm)
Absorption
Emisson
Figure 29: Absorption and emission spectra of PE-Vio 770.
Vio® Dyes technical specifications | April 201714/18
Sample
Conjugate
MFI
Stain
index
Compensation in
channel B2 (%)
A
CD8-PEVio 770
109.2
97.3
0.4
A
CD8-PE-Cy7
67.0
72.7
2.2
B
CD8-PEVio 770
101.0
112.0
0.4
B
CD8-PE-Cy7
65.6
81.7
2.2
Photo-induced conjugate degradation
Analysis of the photo-induced degradation of CD4-PEVio 770 indicated no discernable changes after up to four
hours of continuous exposure to ambient light (~850 Lux).
Significantly higher mean fluorescence intensities (MFI)
and stain indices (SI) for these time points, compare to
commercially available alternatives (BD™ Biosciences),
were also demonstrated (figure 33).
Table 15: MFI and stain indices of CD8-PE-Vio 770 and CD8-PE-Cy7.
Fixation stabilities
Tandem conjugates are usually less stable after fixation
than single absorption/emission fluorochromes. However,
PE-Vio 770 shows excellent stability as shown in figure 32.
Cells fixed and stained with PE-Vio 770, in general, exhibit
between a 0% and 7% loss in fluorescence, making them
excellent candidates for fixation studies.
B
10²
10¹
1
0
-1
10³
CD45RO-FITC
CD45RO-FITC
10³
-1 0 1
10¹
10²
CD45RA-PE-Vio 770
10³
t 120 min
10²
10¹
1
0
-1
-1 0 1
10¹
10²
CD45RA-PE-Vio 770
Figure 32: CD45RA-PE-Vio 770 fluorescence before (A) and after (B)
paraformaldehyde fixation.
10³
CD3-VioGreen
A
t 0 min
t 240 min
CD4
CD4-PE-Vio 770
Marker
MFI
Stain index
Percent spillover
(t0min vs. t 240min )
t 0min t 120min t 240min t 0min t 120min t 240min V1
B2
R1
CD4-PEVio 770
47
46
44
43
42
39
–
0.7/5.5
–
CD4-PECy7
21
21
20
24
24
23
–
0.7/9.0
–
Figure 33: Photo-induced conjugate degradation of PE-Vio 770, with
corresponding MFI and SI. Red lines indicate CD4-PE-Vio 770 (clone
VIT-4.3) from Miltenyi Biotec. Black lines indicate CD4-PE-Cy7 (clone SK3)
from BD Biosciences. Conjugates were exposed to ambient light for up
to four hours, with negligible degradation rates.
Vio® Dyes technical specifications | April 201715/18
Absorption
max. (nm)
Emission
max. (nm)
APC-Vio 770
652
775
APC-Cy7
650
774
APC-Alexa Fluor 750
650
775
Table 16: Absorption and emission maximums of fluorochromes
related to APC-Vio 770.
Laser and filter compatibility
The tandem dyes APC-Vio 770, APC-Cy7, and APC-H7 all
use APC as the donor fluorochrome, showing maximum
absorption at 652 nm. The emission spectra for Vio 770, Cy7,
H7, and Alexa Fluor 750 are similar, making APC-Vio 770 an
ideal tandem conjugate candidate for this channel in all
flow cytometers (figure 34).
100
B 1000
750
750
Side scatter
Fluorochrome
A 1000
500
250
0
-1 0 1
10¹
10²
500
250
0
-1 0 1
10³
CD8-APC-Vio 770
C 1000
D 1000
750
750
500
250
0
-1 0 1
10¹
10²
10¹
10²
10³
CD45-APC-Vio 770
Side scatter
•Large Stokes shift dye, excited with the yellow
(561 nm) or red (635 nm) laser, with an emission
in the near-infrared region at 775 nm
•The combination of Vio® 770 as the acceptor dye and
an optimized chemistry have furnished a tandem dye
that is characterized by a high fluorescence intensity
•Minimal spillover to adjacent detection channels and
low non-specific binding to non-target cells to meet
the complexity of multiparameter cell analysis
•Higher MFI and stain index values, in addition to lower
compensation settings, when compared to APC-Cy7
Side scatter
APC-Vio® 770 Tandem Conjugate
Enhanced brightness
APC-Vio 770 provides strong fluorescent staining,
allowing the identification and analysis of specific cellular
populations (figure 35). When compared to other spectrally
similar conjugates such as APC-Cy7 and APC-H7, APC-Vio 770
exhibits equal or stronger staining patterns (figure 36).
In addition, APC-Vio 770 generally exhibits higher mean
fluorescence intensities (MFI) and greater stain index values,
and requires less compensation in the R1 channel than both
APC-Cy7 and APC-H7 (table 15). These properties make
APC-Vio 770 an ideal fluorochrome for use in the R2 channel.
Side scatter
Red laser dyes
10³
500
250
0
-1 0 1
CD19-APC-Vio 770
10¹
10²
10³
CD4-APC-Vio 770
Figure 35: Human peripheral blood mononuclear cells (PBMC) or
mouse splenocyte (MS) cells were stained with CD8-APC-Vio 770
(A; PBMCs), CD45-APC-Vio 770 (B; PBMCs), CD19-APC-Vio 770 (C; MS),
or CD4-APC-Vio 770 (D; MS) and analyzed by flow cytometry using the
MACSQuant Analyzer.
80
60
APC-Vio770
APC-Vio770
APC-Cy7
APC-Cy7
APC-H7
APC-H7
Relative cell number
Relative cell number
Intensity
70
50
40
30
20
10
0
-1 -1
0 10 1 10¹ 10¹ 10² 10² 10³ 10³
500
550
600
650
700
750
800
850
Wavelength (nm)
APC-Vio 770
Absorption
Emisson
Figure 34: Absorption and emission spectra of APC-Vio 770.
APC-Cy7
APC-H7
Figure 36: Analysis of human PBMCs using CD8 antibodies (clone
BW135/80) conjugated to APC-Vio770, APC-Cy7, or APC-H7. Concurrent
staining with CD14-PerCP and CD56-PE was performed to exclude
CD14 + and CD56+ cells from the analysis.
Vio® Dyes technical specifications | April 201716/18
Sample
Conjugate
MFI
Stain
index
Compensation in
channel R1 (%)
A
CD8-APCVio 770
41.4
52.8
7.0
A
CD8-APC-Cy7
40.6
50.6
11.0
A
CD8-APC-H7
32.2
45.8
9.0
B
CD8-APCVio 770
39.4
57.8
7.0
B
CD8-APC-Cy7
38.8
58.5
11.0
B
CD8-APC-H7
31.2
51.8
9.0
Photo-induced conjugate degradation
Analysis of the photo-induced degradation of
CD8-APC-Vio 770 indicated no discernable changes after
up to 4 hours of continuous exposure to ambient light
(~850 Lux). Significantly higher mean fluorescence
intensities (MFI) and stain indices (SI) for these time points,
compared to commercially available alternatives
(BD Biosciences), were also demonstrated (figure 38).
Table 17: MFI and stain indices of CD8-APC-Vio 770, CD8-APC-Cy7, and
CD8-APC-H7.
t 0 min
Fixation stabilities
APC-Vio® 770 shows excellent stability after fixation with
paraformaldehyde (figure 37), similar to PE-Vio 770. Cellular
material fixed with APC-Vio 770 exhibited, on average, a
fluorescence decrease of between 5% and 10%, indicating
APC-Vio 770’s suitability and stability in fixation experiments.
t 120 min
750
750
500
250
0
-1 0 1
10¹
10²
CD45-APC-Vio 770
10³
500
250
0
-1 0 1
10¹
10²
10³
CD45-APC-Vio 770
Figure 37: Cells were stained with CD45-APC-Vio 770 and left untreated
(A) or fixed with paraformaldehyde (B).
CD3-VioGreen
Side scatter
B 1000
Side scatter
A 1000
t 240 min
CD8
CD8-APC-Vio 770
Marker
MFI
Stain index
Percent spillover
(t 0min vs. t 240 min )
t 0min t 120min t 240min t 0min t 120min t 240min V1
B2
R1
CD8APCVio 770
41
41
39
24
23
22
–
–
9.0/
15.0
CD8APC-Cy7
15
15
15
11
11
11
–
–
9.0/
16.0
Figure 38: Photo-induced conjugate degradation of APC-Vio 770, with
corresponding MFI and SI. Red lines indicate CD8-APC-Vio 770 (clone
BW 135/80) from Miltenyi Biotec. Black lines indicate CD8-APC-Cy7 (clone
RPA-T8) from BD Biosciences. Conjugates were exposed to ambient
light for up to four hours, with negligible degradation rates.
For a complete antibody product list refer to
www.miltenyibiotec.com/antibodies
Vio® Dyes technical specifications | April 201717/18
Germany/Austria/
Switzerland
Miltenyi Biotec GmbH
Friedrich-Ebert-Straße 68
51429 Bergisch Gladbach
Germany
Phone +49 2204 8306-0
Fax +49 2204 85197
[email protected]
USA/Canada
Miltenyi Biotec Inc.
2303 Lindbergh Street
Auburn, CA 95602, USA
Phone 800 FOR MACS
Phone +1 530 888 8871
Fax +1 877 591 1060
[email protected]
Australia
Miltenyi Biotec
Australia Pty. Ltd.
Unit 16 A , 2 Eden Park Drive
Macquarie Park NSW 2113
Australia
Phone +61 2 8877 7400
Fax +61 2 9889 5044
[email protected]
Benelux
Miltenyi Biotec B.V.
Schipholweg 68 H
2316 XE Leiden
The Netherlands
[email protected]
Customer service
The Netherlands
Phone 0800 4020120
Fax 0800 4020100
Customer service Belgium
Phone 0800 94016
Fax 0800 99626
Customer service Luxembourg
Phone 800 24971
Fax 800 24984
France
Miltenyi Biotec SAS
10 rue Mercoeur
75011 Paris, France
Phone +33 1 56 98 16 16
Fax +33 1 56 98 16 17
[email protected]
China
Miltenyi Biotec Technology &
Trading (Shanghai) Co., Ltd.
Rooms 2303 and 2309
No. 319, Xianxia Road
Changning District
200051 Shanghai, P.R. China
Phone +86 21 62351005
Fax +86 21 62350953
[email protected]
Japan
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Nittsu-Eitai Building 5F
16-10 Fuyuki, Koto-ku,
Tokyo 135-0041, Japan
Phone +81 3 5646 8910
Fax +81 3 5646 8911
[email protected]
Italy
Miltenyi Biotec S.r.l.
Via Paolo Nanni Costa, 30
40133 Bologna
Italy
Phone +39 051 6 460 420
Fax +39 051 6 460 499
[email protected]
Nordics and Baltics
Miltenyi Biotec Norden AB
Scheelevägen 17
223 70 Lund
Sweden
[email protected]
Customer service Sweden
Phone 0200-111 800
Fax 046-280 72 99
Customer service Denmark
Phone 80 20 30 10
Fax +46 46 280 72 99
Customer service
Norway, Finland, Iceland,
and Baltic countries
Phone +46 46 280 72 80
Fax +46 46 280 72 99
Singapore
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Phone +65 6238 8183
Fax +65 6238 0302
[email protected]
South Korea
Miltenyi Biotec Korea Co., Ltd
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Phone +82 2 555 1988
Fax +82 2 555 8890
[email protected]
Spain
Miltenyi Biotec S.L.
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Ciudad de la Imagen
28223 Pozuelo de Alarcón (Madrid)
Spain
Phone +34 91 512 12 90
Fax +34 91 512 12 91
[email protected]
United Kingdom
Miltenyi Biotec Ltd.
Almac House, Church Lane
Bisley, Surrey GU24 9DR, UK
Phone +44 1483 799 800
Fax +44 1483 799 811
[email protected]
www.miltenyibiotec.com
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Vio® Dyes technical specifications | April 201718/18